The prostate specific antigen (PSA) was first purified from prostatic tissue (Wang et al. Invest Urol 1979), but the same protein was almost simultaneously and independently characterized in the seminal plasma (Hara et al. J Lab Clin Med 1989; Graves et al. N Engl J Med 1985). PSA is now known to be a 33-kDa glycosylated single chain serine protease (Lilja, J Clin Invest 1985; Watt et al. Proc Natl Acad Sci (U.S.A.) 1986). The 237 amino-acid polyeptide backbone has extensive similarities with that of the glandular kallikreins (Lundwall et al. FEBS Lett 1987; Schaller et al. Eur J Biochem 1987). Unlike the trypsin-like glandular kallikreins, which display Arg-restricted substrate specifity (MacDonald et al. Biochem J 1988), PSA displays chymotrypsin-like substrate specificity (Akiyama et al. FEBS Lett 1987; Christensson et al. Manuscript 1990; Lilja et al. J Biol Chem 1989). PSA has been predicted to be produced as a presumably inactive zymogen (Lundwall et al. FEBS Lett 1987). Active PSA is secreted into the seminal plasma (Lilja, J Clin Invest 1985) where it is one of the most abundant proteins of the prostate (Lilja et al. The Prostate 1988; Dube et al. J Androl 1987). The biological activity of PSA in semen relates to its limited proteolytic fragmentation of the predominant proteins secreted by the seminal vesicles (Lilja, J Clin Invest 1985; Lilja et al. J Clin Invest 1987; McGee et al. Biol Reprod 1988).
Secondary to the release from the prostate epithelium PSA may also be detected in the circulation (Papsidero et al. Cancer Res 1980). Measurements of the serum concentration of PSA have now found widespread use in monitoring of patients with prostate cancer, although increased serum concentrations of PSA have also been reported in benign prostatic hyperplasia and secondary to surgical trauma of the prostate (Duffy, Ann Clin Biochem 1989; Brawer et al. Urology suppl 1989). However, it is presently unknown whether the immunoreactivity in serum represents the PSA-zymogen, the active PSA or PSA inactivated by extracellular proteinase inhibitors and contradictory results have been reported on the molecular mass of this immunoreactivity. Papsidero reported in 1980 the PSA-immunoreactivity to elute as a single 90 to 100 kDa peak (Papsidero et al. Cancer Res 1980), whereas Alfthan and Stenman reported the predominant part of this immunoreactivity to elute as a 30-kDa protein (Alfthan et al. Clin Chem 1988) when subjected to gel filtration chromatography.
In the proceeding invention we showed that PSA has the ability to form complexes with proteinase inhibitors that occur in high concentration in the human extracellular fluids and that PSA occurs in these fluids both in a free and complexed form. In addition, the invention proved to be very useful in diagnosis of prostate cancer patients.